title: Production of homozygous transgenic rainbow trout with enhanced disease resistance.
abstract: Previous studies conducted in our laboratory showed that transgenic medaka expressing cecropin B transgenes exhibited resistant characteristic to fish bacterial pathogens, Pseudomonas fluorescens and Vibrio anguillarum. To confirm whether antimicrobial peptide gene will also exhibit anti-bacterial and anti-viral characteristics in aquaculture important fish species, we produced transgenic rainbow trout expressing cecropin P1 or a synthetic cecropin B analog, CF-17, transgene by sperm-mediated gene transfer method. About 30 % of fish recovered from electroporation were shown to carry the transgene as determined by polymerase chain reaction (PCR) amplification assay. Positive P₁ transgenic fish were crossed to non-transgenic fish to establish F₁ transgenic founder families, and subsequently generating F₂, and F₃ progeny. Expression of cecropin P1 and CF-17 transgenes was detected in transgenic fish by reverse transcription (RT)-PCR analysis. The distribution of body sizes among F₁ transgenic fish were not significantly different from those of non-transgenic fish. Results of challenge studies revealed that many families of F₂ and F₃ transgenic fish exhibited resistance to infection by Aeromonas salmonicida and infectious hematopoietic necrosis virus (IHNV). All-male homozygous cecropin P1 transgenic families were produced by androgenesis from sperm of F₃ heterozygous transgenic fish in one generation. The resistant characteristic to A. salmonicida was confirmed in progeny derived from the outcross of all-male fish to non-transgenic females. Results of our current studies confirmed the possibility of producing disease-resistant homozygous rainbow trout strains by transgenesis of cecropin P1 or CF-17 gene and followed by androgenesis.
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Road, U-3125, Storrs, CT, 06269, USA.
authors.forename: Pinwen Peter
authors.initials: PP
authors.lastname: Chiou
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Chen
authors.forename: Chun-Mean
authors.initials: CM
authors.lastname: Lin
authors.forename: Jenny
authors.initials: J
authors.lastname: Khoo
authors.forename: Jon
authors.initials: J
authors.lastname: Larson
authors.forename: Rich
authors.initials: R
authors.lastname: Holt
authors.forename: Jo-Ann
authors.initials: JA
authors.lastname: Leong
authors.forename: Gary
authors.initials: G
authors.lastname: Thorgarrd
authors.forename: Thomas T
authors.initials: TT
authors.lastname: Chen
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D017756:Aquaculture / Q000379:methods; D003433:Crosses, Genetic; D017931:DNA Primers / Q000235:genetics; D060467:Disease Resistance / Q000235:genetics*; D018274:Electroporation / Q000662:veterinary; D005260:Female; D018014:Gene Transfer Techniques / Q000662:veterinary; D006801:Humans; D008297:Male; D017686:Oncorhynchus mykiss / Q000235:genetics*; D010455:Peptides / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
chemical_list: D017931:DNA Primers; D010455:Peptides; C070016:cecropin P1-LI